Abstract:

A separation method based on high-speed counter-current chromatography (HSCCC) has been established for the isolation and preparation of gallic acid from the ethanol extract of Terminalia chebula Retz. After comparing five kinds of solvent protocols of HSCCC, the two-phase system of n-hexane-ethyl acetate-methanol-water (1:5:1:5, v/v/v/v) was finally chosen as the operating solvent of HSCCC for the separation of gallic acid, in which the lower phase was used as the mobile phase and the upper phase as stationary phase. The detection in the experiments was performed with an ultraviolet detector at 254 nm. Under the conditions of rotation speed of 850 r/min, lower phase flow rate of 2 mL/min, four peaks (Ⅰ/Ⅱ/Ⅲ/Ⅳ) were displayed on HSCCC chromatogram. Among them, only peak Ⅲ contained a large amount of gallic acid (about 96.40%), which was confirmed by electrospray ionization mass spectrometry (ESI-MS) and high performance liquid chromatographic (HPLC) analysis. As much as 8.6 mg of gallic acid was obtained from 100 mg of the ethanol extract of Terminalia chebula Retz., indicating the method is simple, robust and efficient for the preparation of gallic acid.

Key words: Terminalia chebula Retz, gallic acid, high-speed counter-current chromatography (HSCCC)